CCL2/MCP1 Antibody - BSA Free

Western Blot: CCL2/MCP1 Antibody [NBP1-07035] - Western blot for MCP-1 protein was analyzed from the ox-LDL-stimulated THP-1 cells. B-actin was used as loading control (D-I). Scale bar: 20 um. Bar graph indicates the mean +/- SD (n = 3). *P < 0.05 and **P < 0.01 vs. Cont group (0 ug/mL of ox-LDL). Image collected and cropped by Citeab from the following publication (SIRT1 inhibition promotes atherosclerosis through impaired autophagy. Oncotarget (2017)) licensed under a CC-BY license.

Immunohistochemistry-Paraffin: CCL2/MCP1 Antibody [NBP1-07035] - IHC analysis of a formalin-fixed paraffin-embedded (FFPE) human breast carcinoma tissue section using 1:1000 dilution of CCL2/MCP1 antibody (NBP1-07035) on a Bond Rx autostainer (Leica Biosystems). The assay involved 20 minutes of heat induced antigen retrieval (HIER) with 10mM sodium citrate buffer (pH 6.0) and endogenous peroxidase quenching using peroxide block. The sections were incubated with primary antibody for 30 minutes. Bond Polymer Refine Detection (Leica Biosystems) and DAB were used for signal detection which followed counterstaining with hematoxylin. Whole slide scanning and capturing of representative images (20X) were performed using Aperio AT2 (Leica Biosystems). This antibody generated a diffused cytoplasmic staining of CCL2 antigen in the cancer cells, stromal cells as well as the endothelial cells. The stroma itself showed a weak immunopositivity for CCL2. Staining was performed by Histowiz.

Western Blot: CCL2/MCP1 Antibody [NBP1-07035] - Inhibition of SIRT1 using EX-527 or SIRT1 siRNA transfection enhanced MCP-1 expression and foam cell formationHuman THP-1 macrophages were pretreated with EX-527 (2 uM, for 2 hrs) or SIRT1 siRNA (20 uM, for 24 hrs), and then exposed to 80 i1/4g/mL of ox-LDL for an additional 24 hrs. Western blot for SIRT1 and MCP-1 proteins were analyzed from the ox-LDL-stimulated THP-1 cells. I2-actin was used as loading control. Image collected and cropped by Citeab from the following publication (SIRT1 inhibition promotes atherosclerosis through impaired autophagy. Oncotarget (2017)) licensed under a CC-BY license.

Immunocytochemistry/Immunofluorescence: CCL2/MCP1 Antibody [NBP1-07035] - MCP1 antibody was tested in HeLa cells with Dylight 488 (green). Nuclei and alpha-tubulin were counterstained with DAPI (blue) and Dylight 550 (red).

Immunohistochemistry-Paraffin: CCL2/MCP1 Antibody [NBP1-07035] - IHC analysis of a formalin-fixed paraffin-embedded (FFPE) human breast carcinoma tissue section using 1:1000 dilution of CCL2/MCP1 antibody (NBP1-07035) on a Bond Rx autostainer (Leica Biosystems). The assay involved 20 minutes of heat induced antigen retrieval (HIER) with 10mM sodium citrate buffer (pH 6.0) and endogenous peroxidase quenching using peroxide block. The sections were incubated with primary antibody for 30 minutes. Bond Polymer Refine Detection (Leica Biosystems) and DAB were used for signal detection which followed counterstaining with hematoxylin. Whole slide scanning and capturing of representative images (20X) were performed using Aperio AT2 (Leica Biosystems). This antibody generated a diffused cytoplasmic staining of CCL2 antigen in the cancer cells, stromal cells as well as the endothelial cells. The stroma itself showed a weak immunopositivity for CCL2. Staining was performed by Histowiz.

Western Blot: CCL2/MCP1 Antibody [NBP1-07035] - CCL2 expression level was evaluated in control group and aLAG-3 group.

Western Blot: CCL2/MCP1 Antibody - BSA Free [NBP1-07035] - Ox-LDL induced macrophage foam cell formation, SIRT1 inhibition, autophagy impairment, & MCP-1 production in THP-1 cellsHuman THP-1 macrophages were exposed to 0, 20, 40, 60, & 80 μg/mL of ox-LDL for 24 hrs. Treated cells were photographed using light microscopy (A). The THP-1 macrophage-derived foam cell formation was determined using ORO staining method (B) & (C). Western blot for SIRT1, LC3, Beclin1, p62, & MCP-1 proteins were analyzed from the ox-LDL-stimulated THP-1 cells. beta -actin was used as loading control (D-I). Scale bar: 20 μm. Bar graph indicates the mean ± SD (n = 3). *P < 0.05 & **P < 0.01 vs. Cont group (0 μg/mL of ox-LDL). Image collected & cropped by CiteAb from the following publication (//www.oncotarget.com/lookup/doi/10.18632/oncotarget.17691), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: CCL2/MCP1 Antibody - BSA Free [NBP1-07035] - Inhibition of autophagy with 3-MA led to increased foam cell formation & MCP-1 productionHuman THP-1 macrophages were pretreated with 3-MA (5 μM) for 2 hrs, & then exposed to 80 μg/mL of ox-LDL for an additional 24 hrs. Western blot for LC3, Beclin1, p62, & MCP-1 proteins were analyzed from the ox-LDL-stimulated THP-1 cells. beta -actin was used as loading control (A-E). THP-1 macrophage-derived foam cell formation was determined using ORO staining method (F-G). Scale bar: 40 μm. Bar graph indicates the mean ± SD (n = 3). *P < 0.05 & **P < 0.01 vs. Cont group; #P < 0.05 & ##P < 0.01 vs. 3-MA group; &P < 0.05 & &&P < 0.01 represent significant differences between ox-LDL group & 3-MA+ox-LDL group. Image collected & cropped by CiteAb from the following publication (//www.oncotarget.com/lookup/doi/10.18632/oncotarget.17691), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: CCL2/MCP1 Antibody - BSA Free [NBP1-07035] - CCL2-ir cellular distribution in ependymal layers attached to the CC. A, B CCL2-ir (CCL2 panels, arrows) & GFAP (GFAP panels, arrowheads) double immunostained structures (Merged panels, arrow-arrowheads) are seen inlaid in the ependymal layers attached to the CC in both saline & LPS injected mice. C A CCL2-ir & Iba-1 double labeled soma (Merged, arrow-arrowhead) is seen at a corner of lateral ventricle, which seems be just next to an invaginated choroid plexus between the CC & basal ganglion (framed areas & insets). D, E Hyper-ramified & amoeba like Iba-1 labeled cells (Iba-1 panel, opened arrowheads) are observed in the CC of LPS injected mice, indicating their activated states. The CCL2-ir & Iba-1 double labeled structures are also regarded in the ependymal layers (Merged, arrow-arrowheads). Scare bar = 100 µm in all A– E Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/35354428), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: CCL2/MCP1 Antibody - BSA Free [NBP1-07035] - Inhibition of autophagy using Atg5 siRNA aggravated foam cell formation & MCP-1 expressionHuman THP-1 macrophages were pretreated with Atg5 siRNA (20 μM) for 24 hrs, & then exposed to 80 μg/mL of ox-LDL for an additional 24 hrs. Western blot forAtg5, LC3, Beclin1, p62, & MCP-1 proteins were analyzed from the ox-LDL-stimulated THP-1 cells. beta -actin was used as loading control (A-F) & (I). THP-1 macrophage-derived foam cell formation was determined using ORO staining method (G) & (H). Scale bar: 40 μm. Bar graph indicates the mean ± SD (n = 3). *P < 0.05 & **P < 0.01 vs. NC siRNA group; #P < 0.05 & ##P < 0.01 vs. Atg5 siRNA group; &P < 0.05 & &&P < 0.01 represent significant differences between NC siRNA+ox-LDL group & Atg5 siRNA+ox-LDL group. Image collected & cropped by CiteAb from the following publication (//www.oncotarget.com/lookup/doi/10.18632/oncotarget.17691), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: CCL2/MCP1 Antibody - BSA Free [NBP1-07035] - Inhibition of SIRT1 using EX-527 or SIRT1 siRNA transfection enhanced MCP-1 expression & foam cell formationHuman THP-1 macrophages were pretreated with EX-527 (2 μM, for 2 hrs) or SIRT1 siRNA (20 μM, for 24 hrs), & then exposed to 80 μg/mL of ox-LDL for an additional 24 hrs. Western blot for SIRT1 & MCP-1 proteins were analyzed from the ox-LDL-stimulated THP-1 cells. beta -actin was used as loading control (A-C) & (F-H). THP-1 macrophage-derived foam cell formation was determined using ORO staining method (D-E) & (I-J). Scale bar: 40 μm. Bar graph indicates the mean ± SD (n = 3). *P < 0.05 & **P < 0.01 vs. Cont group (NC siRNA group); #P < 0.05 & ##P < 0.01 vs. EX-527 group (SIRT1 siRNA group); &P < 0.05 & &&P < 0.01 represent significant differences between ox-LDL group (NC siRNA+ox-LDL group) & EX-527+ox-LDL group (SIRT1 siRNA+ox-LDL group). Image collected & cropped by CiteAb from the following publication (//www.oncotarget.com/lookup/doi/10.18632/oncotarget.17691), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: CCL2/MCP1 Antibody - BSA Free [NBP1-07035] - Overexpression of SIRT1 using adenoviral transfection reversed ox-LDL-induced macrophage foam cell formation & autophagy impairment in THP-1 cellsHuman THP-1 macrophages were transfected by SIRT1 over-expressing adenovirus (HBAD-SIRT1) or NC adenovirus (HBAD-GFP) for 24 hrs & then exposed to 80 μg/mL of ox-LDL for an additional 24 hrs. The transfected THP-1 cells were observed using an inverted fluorescence microscope (A) & then were harvested for transfection efficiency analysis by Western blot method (B). THP-1 macrophage-derived foam cell formation was determined using ORO staining method (C) & (D). Western blot for LC3, Beclin1, p62, & Atg5 proteins & immunoprecipitation for acetyl-Lys Atg5 were analyzed from the ox-LDL-stimulated THP-1 cells. beta -actin was used as loading control (E-L). Scale bar: 40 μm. Bar graph indicates the mean ± SD (n = 3). *P < 0.05 & **P < 0.01 vs. HBAD-GFP group; #P < 0.05 & ##P < 0.01 vs. HBAD-SIRT1 group; &P < 0.05 & &&P < 0.01 represent significant differences between HBAD-GFP+ox-LDL group & HBAD-SIRT1+ox-LDL group. Image collected & cropped by CiteAb from the following publication (//www.oncotarget.com/lookup/doi/10.18632/oncotarget.17691), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

• Reactivity: Hu, Mu, Rt
• Applications: WB, ICC/IF, IHC, B/N
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Format: BSA Free
• Concentration: 1 mg/ml

CCL2/MCP1 Antibody - BSA Free Summary

• Immunogen: Synthetic peptide made to an internal portion of rat MCP1 (within residues 15-40). [Swiss-Prot# P14844]
• Localization: Secreted, secretory vesicles
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: CCL2
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • Block/Neutralize reported in scientific literature (PMID 22778093)
  • Immunocytochemistry/ Immunofluorescence 1:2000
  • Immunohistochemistry 1:500-1:1000
  • Immunohistochemistry-Paraffin 1:500-1:1000
  • Western Blot 1:1000. Use reported in scientific literature (PMID 22402584)
• Application Notes: In Western blot, a band is seen at ~16 kDa. In ICC/IF secretory vesicles staining was observed in HeLa cells.
• Theoretical MW: 16 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• Control:

  CCL2/MCP1 Overexpression Lysate

• Control Peptide:

  CCL2/MCP1 Peptide (NBP1-07035PEP)

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: 0.02% Sodium Azide
• Concentration: 1 mg/ml
• Purity: Immunogen affinity purified

Alternate Names for CCL2/MCP1 Antibody - BSA Free

• C-C motif chemokine 2
• chemokine (C-C motif) ligand 2
• GDCF-2
• HC11Monocyte secretory protein JE
• MCAFMonocyte chemotactic and activating factor
• MCP1 Monocyte chemotactic protein 1
• MCP-1 Small-inducible cytokine A2
• MGC9434
• Monocyte chemoattractant protein 1
• monocyte chemoattractant protein-1
• SCYA2HSMCR30
• small inducible cytokine A2 (monocyte chemotactic protein 1, homologous tomouse Sig-je)
• small inducible cytokine subfamily A (Cys-Cys), member 2
• SMC-CF

Background

MCP-1 (monocyte chemoattracctant protein-1, also called C-C motif chemokine 2 or CCL2) is a secreted protein belonging to the intercrine beta (chemokine CC) family and specifically acts as a chemotactic factor to attract monocytes, CD4+ T cells, NK cells, and CCR2 expressing cells, but not neutrophils. Both hematopoietic and non-hematopoietic cells that include monocytes/macrophages, dendritic cells, astrocytes, endothelial cells and fibroblasts express MCP-1, and monocytes/macrophages are the major source of MCP-1 in peripheral blood. In addition to expression at acute inflammation sites, MCP-1 is upregulated in chronic inflammation, including autoimmune disease, atherosclerosis, cancer, and chronic infection. MCP-1 has been implicated in both pro-inflammatory and anti-inflammatory responses, and is a potential intervention point for the treatment of various inflammatory/autoimmune diseases (multiple sclerosis, atherosclerosis, insulin-resistant diabetes, rheumatoid arthritis and lupus nephritis), cancers (breast and prostate cancer) and HIV-1/AIDS.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for CCL2/MCP1 Antibody (NBP1-07035)(21)

Publications using NBP1-07035

• Zhao H, Liu Z, Chen H et al. Identifying specific functional roles for senescence across cell types. Cell 2024-10-01 [PMID: 39368477]
• Zhou Jiang, Qianpei He, Jackson Wezeman, Martin Darvas, Warren Ladiges A cocktail of rapamycin, acarbose, and phenylbutyrate prevents age-related cognitive decline in mice by targeting multiple aging pathways. GeroScience 2024-05-16 [PMID: 38755466]
• T. Ollewagen, R.M. Benecke, C. Smith High species homology potentiates quantitative inflammation profiling in zebrafish using immunofluorescence Heliyon 2023-12-13 [PMID: 38187273]
• Deng S, Zhou F, Wang F et al. C5a enhances V?1 T cells recruitment via the CCL2-CCR2 axis in IgA nephropathy International immunopharmacology 2023-10-18 [PMID: 37862725] (Human)
  
  Details:
  HMC Stimulation
• Wilkinson A, Hulme S, Kennedy J et al. The senescent secretome drives PLVAP expression in cultured human hepatic endothelial cells to promote monocyte transmigration iScience 2023-09-01 [PMID: 37810232]
• Andre AB, Rees KP, O'Connor S et al. Single cell analysis reveals satellite cell heterogeneity for proinflammatory chemokine expression Frontiers in cell and developmental biology 2023-03-27 [PMID: 37051469] (IHC-P, Mouse)
  
  Details:
  1:500 IHC-P dilution
• Zhu X, Schrader JM, Irizarry BA et al. Impact of A beta 40 and A beta 42 Fibrils on the Transcriptome of Primary Astrocytes and Microglia Biomedicines 2022-11-19 [PMID: 36428550] (IHC-Fr, Rat)
  
  Details:
  Fifteen-month-old rTgF344-AD rat brain sections, dilution used 1:200
• Shi, X;Gong, X;Xiong, H;Zhang, J; Cellular distribution of C-C motif chemokine ligand 2 like immunoreactivities in frontal cortex and corpus callosum of normal and lipopolysaccharide treated animal BMC Neuroscience [PMID: 35354428] (ICC/IF)
• Ollewagen T, Powrie YSL, Myburgh KH, Smith C Unresolved intramuscular inflammation, not diminished skeletal muscle regenerative capacity, is at the root of rheumatoid cachexia: insights from a rat CIA model Physiological reports 2021-11-01 [PMID: 34806343] (WB, Rat)
• Yitong Zhao, Masaki Goto, Nosratola D Vaziri, Mahyar Khazaeli, Han Liu, Nazli Farahanchi, Elham Khanifar, Ted Farzaneh, Patrick A Haslett, Hamid Moradi, Mangala M Soundarapandian RNA Interference Targeting Liver Angiopoietin-Like Protein 3 Protects from Nephrotic Syndrome in a Rat Model Via Amelioration of Pathologic Hypertriglyceridemia. The Journal of pharmacology and experimental therapeutics 2021-04-27 [PMID: 33443084]
• Zheng Y, Lu P, Deng Y et al. Single-Cell Transcriptomics Reveal Immune Mechanisms of the Onset and Progression of IgA Nephropathy Cell reports 2020-12-22 [PMID: 33357427] (ICC/IF, Human)
• Wu J, Dou Y, Ladiges W Adverse neurological effects of short-term sleep deprivation in aging mice are prevented by SS31 peptide Clocks Sleep 2020-10-22 [PMID: 33089207]
• Zhao L, Fan M, Zhao L et al. Fibroblast growth factor 1 ameliorates adipose tissue inflammation and systemic insulin resistance via enhancing adipocyte mTORC2/Rictor signal J. Cell. Mol. Med. 2020-09-26 [PMID: 32979037] (IHC-P, WB, Mouse)
• Chen I-Chen, Lin Yu-Tsai, Huang Jhy-Shrian et al. Decreased Ambient Oxygen Tension Alters the Expression of Endothelin-1, iNOS and cGMP in Rat Alveolar Macrophages. International Journal of Medical Sciences 2019-02-28 [PMID: 30911278] (WB, Rat)
• Alghamdi TA, Batchu SN, Hadden MJ, et al. Histone H3 Serine 10 Phosphorylation Facilitates Endothelial Activation in Diabetic Kidney Disease Diabetes 2018-12-01 [PMID: 30213824] (WB, Mouse)
• Yang X, Wei J, He Y et al. SIRT1 inhibition promotes atherosclerosis through impaired autophagy Oncotarget 2017-08-01 [PMID: 28881659] (ICC/IF, Human)
• Verzola D, Milanesi S, Bertolotto M et al. Myostatin mediates abdominal aortic atherosclerosis progression by inducing vascular smooth muscle cell dysfunction and monocyte recruitment. Scientific Reports 2017-01-01 [PMID: 28406165] (WB, ICC/IF, Human)
• Deng W, Mao L, Yu G et al. LAG-3 confers poor prognosis and its blockade reshapes antitumor response in head and neck squamous cell carcinoma. Oncoimmunology 2016-10-07 [PMID: 27999760] (WB, Mouse)
• Choi JA, Chung YR, Byun HR et al. The anti-ALS drug riluzole attenuates pericyte loss in the diabetic retinopathy of streptozotocin-treated mice. Toxicol Appl Pharmacol 2017-01-15 [PMID: 27939241] (Mouse)
• Lamont EA, O'Grady SM, Davis WC et al. Infection with Mycobacterium avium subsp. paratuberculosis Results in Rapid Interleukin-1 beta Release and Macrophage Transepithelial Migration Infect Immun 2012-09-01 [PMID: 22778093] (B/N, Human)
• Fet NG, Fiebeler A, Klinge U et al. Reduction of activated macrophages after ischaemia-reperfusion injury diminishes oxidative stress and ameliorates renal damage Nephrol Dial Transplant 2012-03-08 [PMID: 22402584] (IF/IHC, Rat)

Review for CCL2/MCP1 Antibody (NBP1-07035) (1)

Average Rating: 5

(Based on 1 review)

We have 1 review tested in 1 species: Mouse.

reviewed by: Verified Customer WB Mouse 10/23/2017

Summary

• Application: Western Blot
• Sample Tested: Mouse Head and neck squamous cell carcinoma (HNSCC)
• Species: Mouse
• Lot: NBP1-07035

Bioinformatics

• Gene Symbol: CCL2